生物多样性 ›› 2017, Vol. 25 ›› Issue (6): 627-637. DOI: 10.17520/biods.2017090
郑硕理1,2, 田晓玲3, 黄承玲3, 王灵军4, 冯元4, 张敬丽1,*()
收稿日期:
2017-03-22
接受日期:
2017-06-28
出版日期:
2017-06-20
发布日期:
2017-07-10
通讯作者:
张敬丽
基金资助:
Shuoli Zheng1,2, Xiaoling Tian3, Chengling Huang3, Lingjun Wang4, Yuan Feng4, Jingli Zhang1,*()
Received:
2017-03-22
Accepted:
2017-06-28
Online:
2017-06-20
Published:
2017-07-10
Contact:
Zhang Jingli
摘要:
中国西南地区是世界杜鹃属(Rhododendron)植物的分布和分化中心, 开展杜鹃属自然杂交研究可为分类修订和新品种选育提供依据。本文以贵州百里杜鹃自然保护区(百里保护区)和云南石宝山分布的大白杜鹃(Rhododendron decorum)、马缨杜鹃(R. delavayi)及两种间疑似自然杂交后代和人工杂交后代个体为研究材料, 通过形态特征比较、核基因间隔序列ITS以及叶绿体基因片段trnL-trnF、rbcL和trnH-psbA测序特征分析, 发现疑似自然杂交后代与人工杂交后代形态特征介于大白杜鹃与马缨杜鹃之间; 4个DNA片段序列均能区分大白杜鹃和马缨杜鹃, 同一物种的不同居群在叶绿体基因片段rbcL、trnH-psbA序列存在差异位点; 所有人工杂交后代个体和1个石宝山疑似杂交个体均以大白杜鹃为母本, 所有百里保护区疑似杂交个体和6个石宝山疑似杂交个体以马缨杜鹃为母本。本研究证实了大白杜鹃与马缨杜鹃在百里保护区和云南石宝山存在自然杂交现象, 杂交为双向且自然杂交后代绝大多数以马缨杜鹃为母本。
郑硕理, 田晓玲, 黄承玲, 王灵军, 冯元, 张敬丽 (2017) 结合分子手段和形态分析验证大白杜鹃与马缨杜鹃的自然杂交. 生物多样性, 25, 627-637. DOI: 10.17520/biods.2017090.
Shuoli Zheng, Xiaoling Tian, Chengling Huang, Lingjun Wang, Yuan Feng, Jingli Zhang (2017) Molecular and morphological evidence for natural hybridization between Rhododendron decorum and R. delavayi (Ericaceae). Biodiversity Science, 25, 627-637. DOI: 10.17520/biods.2017090.
编号 Code | 种类 Taxon | 地点 Site | 经纬度 Latitude and longitude | 海拔 Altitude (m) | 个体数量 Sample size |
---|---|---|---|---|---|
BLM (1-10) | 马缨杜鹃 R. delavayi | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 10 |
BLD (1-10) | 大白杜鹃 R. decorum | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 10 |
BLH (1-8) | 疑似杂交后代 Putative hybrids | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 8 |
SBSM (1-5) | 马缨杜鹃 R. delavayi | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 5 |
SBSD (1-5) | 大白杜鹃 R. decorum | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 5 |
SBSH (2/3/4/5/7/8/9) | 疑似杂交后代 Putative hybrids | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 7 |
AH (1-5) | 人工杂交后代 Artificial hybrids | 昆明植物园 Kunming Botanical Garden | 5 | ||
总计 Total | 50 |
表1 大白杜鹃、马缨杜鹃及疑似杂交后代的来源和数量
Table 1 Sources and sample size of Rhododendron decorum, R. delavayi, and putative hybrids used in this study
编号 Code | 种类 Taxon | 地点 Site | 经纬度 Latitude and longitude | 海拔 Altitude (m) | 个体数量 Sample size |
---|---|---|---|---|---|
BLM (1-10) | 马缨杜鹃 R. delavayi | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 10 |
BLD (1-10) | 大白杜鹃 R. decorum | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 10 |
BLH (1-8) | 疑似杂交后代 Putative hybrids | 贵州百里杜鹃自然保护区 Baili Rhododendron Nature Reserve in Guizhou Province | 27°15′ N 105°58′ E | 1,700-1,800 | 8 |
SBSM (1-5) | 马缨杜鹃 R. delavayi | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 5 |
SBSD (1-5) | 大白杜鹃 R. decorum | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 5 |
SBSH (2/3/4/5/7/8/9) | 疑似杂交后代 Putative hybrids | 云南大理石宝山 Shibao Mountain of Dali, Yunnan | 26°22′ N 99°50′ E | 2,400-2,600 | 7 |
AH (1-5) | 人工杂交后代 Artificial hybrids | 昆明植物园 Kunming Botanical Garden | 5 | ||
总计 Total | 50 |
形态特征 Morphological characters | 大白杜鹃 R. decorum | 疑似自然杂交与人工杂交后代 Putative and artificial hybrids | 马缨杜鹃 R. delavayi |
---|---|---|---|
花冠颜色 Corolla color | 白色 White | 粉红 Pink | 红色 Red |
花冠裂片数 No. of corolla lobes | 6-8, 通常7 6-8, usually 7 | 5-6, 无规律 5-6, irregular | 5 |
叶片形状 Leaf shape | 长圆形 Oblong | 长圆形至披针形 Oblong to lanceolate | 披针形 Lanceolate |
叶片毛被 Ventral leaf suraface indumentum | 无毛 Glabrous | 稀疏毛被 Thin indumentum | 浓密海绵状毛被 Woolly indumentum |
果实表面特征 Fruit surface character | 有腺体 Glands | 有腺体或毛被或两者均有 Glands or/and indumentum | 密被棕红色毛被 Continuous brownish red indumentum |
花期 Flowering period | 5-6月 May to June | 4月中旬-5月中旬 Mid-April to mid-May | 3月-5月 March to May |
表2 大白杜鹃、马缨杜鹃、人工杂交后代与疑似杂交后代的形态特征对比
Table 2 Comparison of morphological characters of Rhododendron decorum, R. delavayi, artificial and putative hybrids
形态特征 Morphological characters | 大白杜鹃 R. decorum | 疑似自然杂交与人工杂交后代 Putative and artificial hybrids | 马缨杜鹃 R. delavayi |
---|---|---|---|
花冠颜色 Corolla color | 白色 White | 粉红 Pink | 红色 Red |
花冠裂片数 No. of corolla lobes | 6-8, 通常7 6-8, usually 7 | 5-6, 无规律 5-6, irregular | 5 |
叶片形状 Leaf shape | 长圆形 Oblong | 长圆形至披针形 Oblong to lanceolate | 披针形 Lanceolate |
叶片毛被 Ventral leaf suraface indumentum | 无毛 Glabrous | 稀疏毛被 Thin indumentum | 浓密海绵状毛被 Woolly indumentum |
果实表面特征 Fruit surface character | 有腺体 Glands | 有腺体或毛被或两者均有 Glands or/and indumentum | 密被棕红色毛被 Continuous brownish red indumentum |
花期 Flowering period | 5-6月 May to June | 4月中旬-5月中旬 Mid-April to mid-May | 3月-5月 March to May |
编号 Code | 克隆数量 Clone numbers | 类型 Type | 差异位点 Variable sites (bp) | 类型 Type | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|
94 | 101 | 114 | 203 | 494 | 505 | A1 | A2 | A4 | |||
大白杜鹃 R. decorum | 0 | A1 | T | G | G | T | T | C | 15 | 0 | 0 |
马缨杜鹃 R. delavayi | 0 | A2 | C | C | T | G | C | T | 0 | 15 | 0 |
AH、BLH、SBSH | A3 | Y | S | K | K | Y | Y | ||||
克隆测序 Clone sequences | |||||||||||
BLH1 | 2 | 1 | 1 | 0 | |||||||
BLH2 | 4 | 1 | 2 | 1 | |||||||
BLH3 | 3 | 2 | 1 | 0 | |||||||
BLH4 | 3 | 2 | 1 | 0 | |||||||
BLH5 | 3 | 1 | 1 | 1 | |||||||
BLH6 | 5 | 1 | 1 | 3 | |||||||
BLH7 | 4 | 2 | 1 | 1 | |||||||
BLH8 | 4 | 2 | 1 | 1 | |||||||
AH1 | 3 | 1 | 2 | 1 | |||||||
AH2 | 3 | 1 | 2 | 0 | |||||||
AH3 | 4 | 2 | 2 | 0 | |||||||
AH4 | 3 | 2 | 1 | 0 | |||||||
AH5 | 4 | 2 | 2 | 1 | |||||||
SBSH2 | 6 | 4 | 2 | 0 | |||||||
SBSH3 | 8 | 5 | 3 | 0 | |||||||
SBSH4 | 5 | 4 | 1 | 0 | |||||||
SBSH5 | 5 | 4 | 1 | 0 | |||||||
SBSH7 | 7 | 5 | 2 | 0 | |||||||
SBSH8 | 7 | 6 | 1 | 0 | |||||||
SBSH9 | 5 | 3 | 2 | 0 | |||||||
克隆数量总计 Total clone numbers | 53 | 31 | 10 |
表3 大白杜鹃与马缨杜鹃的ITS序列差异位点及疑似杂交后代克隆序列
Table 3 The location of variable sites based on ITS and clone sequences of Rhododendron decorum, R. delavayi and putative hybrids
编号 Code | 克隆数量 Clone numbers | 类型 Type | 差异位点 Variable sites (bp) | 类型 Type | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|
94 | 101 | 114 | 203 | 494 | 505 | A1 | A2 | A4 | |||
大白杜鹃 R. decorum | 0 | A1 | T | G | G | T | T | C | 15 | 0 | 0 |
马缨杜鹃 R. delavayi | 0 | A2 | C | C | T | G | C | T | 0 | 15 | 0 |
AH、BLH、SBSH | A3 | Y | S | K | K | Y | Y | ||||
克隆测序 Clone sequences | |||||||||||
BLH1 | 2 | 1 | 1 | 0 | |||||||
BLH2 | 4 | 1 | 2 | 1 | |||||||
BLH3 | 3 | 2 | 1 | 0 | |||||||
BLH4 | 3 | 2 | 1 | 0 | |||||||
BLH5 | 3 | 1 | 1 | 1 | |||||||
BLH6 | 5 | 1 | 1 | 3 | |||||||
BLH7 | 4 | 2 | 1 | 1 | |||||||
BLH8 | 4 | 2 | 1 | 1 | |||||||
AH1 | 3 | 1 | 2 | 1 | |||||||
AH2 | 3 | 1 | 2 | 0 | |||||||
AH3 | 4 | 2 | 2 | 0 | |||||||
AH4 | 3 | 2 | 1 | 0 | |||||||
AH5 | 4 | 2 | 2 | 1 | |||||||
SBSH2 | 6 | 4 | 2 | 0 | |||||||
SBSH3 | 8 | 5 | 3 | 0 | |||||||
SBSH4 | 5 | 4 | 1 | 0 | |||||||
SBSH5 | 5 | 4 | 1 | 0 | |||||||
SBSH7 | 7 | 5 | 2 | 0 | |||||||
SBSH8 | 7 | 6 | 1 | 0 | |||||||
SBSH9 | 5 | 3 | 2 | 0 | |||||||
克隆数量总计 Total clone numbers | 53 | 31 | 10 |
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
147 | 622 | 623 | 624 | 626 | 627 | 629 | 646 | 831 | ||
大白杜鹃 Rhododendron decorum | B1 | G | T | T | T | T | T | T | C | T |
马缨杜鹃 R. delavayi | B2 | T | A | A | A | A | A | A | A | C |
AH、SBSH5 | B1 | G | T | T | T | T | T | T | C | T |
BLH、SHSH2/3/4/7/8/9 | B2 | T | A | A | A | A | A | A | A | C |
表4 大白杜鹃、马缨杜鹃及疑似杂交后代的trnL-trnF序列差异位点
Table 4 The location of variable sites based on trnL-trnF sequences of Rhododendron decorum, R. delavayi and putative hybrids
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
147 | 622 | 623 | 624 | 626 | 627 | 629 | 646 | 831 | ||
大白杜鹃 Rhododendron decorum | B1 | G | T | T | T | T | T | T | C | T |
马缨杜鹃 R. delavayi | B2 | T | A | A | A | A | A | A | A | C |
AH、SBSH5 | B1 | G | T | T | T | T | T | T | C | T |
BLH、SHSH2/3/4/7/8/9 | B2 | T | A | A | A | A | A | A | A | C |
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||
---|---|---|---|---|---|---|
8 | 30 | 263 | 398 | 407 | ||
BLD | C1 | C | G | T | C | C |
BLM | C2 | C | T | A | T | T |
SBSD | C3 | G | G | T | C | C |
SBSM | C4 | G | T | A | T | T |
AH | C1 | C | G | T | C | C |
BLH | C2 | C | T | A | T | T |
SBSH5 | C3 | G | G | T | C | C |
SBSH2/3/4/7/8/9 | C4 | G | T | A | T | T |
表5 大白杜鹃、马缨杜鹃及疑似杂交后代的rbcL序列差异位点
Table 5 The location of variable sites based on rbcL sequences of Rhododendron decorum, R. delavayi and putative hybrids
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||
---|---|---|---|---|---|---|
8 | 30 | 263 | 398 | 407 | ||
BLD | C1 | C | G | T | C | C |
BLM | C2 | C | T | A | T | T |
SBSD | C3 | G | G | T | C | C |
SBSM | C4 | G | T | A | T | T |
AH | C1 | C | G | T | C | C |
BLH | C2 | C | T | A | T | T |
SBSH5 | C3 | G | G | T | C | C |
SBSH2/3/4/7/8/9 | C4 | G | T | A | T | T |
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 128 | 135 | 211 | 214 | 245 | 254 | 254 | 299 | 313 | 333 | 337 | 340 | 344 | 348 | 351 | 358 | 365-369 | ||
BLD | D1 | G | A | T | C | G | C | G | A | C | A | A | G | T | G | G | T | T | T | AATTC |
SBSD | D2 | - | G | * | * | * | * | * | * | * | * | * | A | A | A | A | A | G | G | ***** |
BLM | D3 | * | * | A | G | A | C | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
SBSM | D4 | * | * | A | G | A | A | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
AH | D1 | G | A | T | C | G | C | G | A | C | A | A | G | T | G | G | T | T | T | AATTC |
SBSH5 | D2 | * | G | * | * | * | * | * | * | * | * | * | A | A | A | A | A | G | G | ***** |
BLH | D3 | * | * | A | G | A | C | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
SBSH2/3/4/7/8/9 | D4 | * | * | A | G | A | A | T | T | T | T | G | * | * | * | * | * | * | · | G---- |
表6 大白杜鹃、马缨杜鹃及疑似杂交后代的trnH-psbA序列差异位点
Table 6 The location of variable sites based on trnH-psbA sequences of Rhododendron decorum, R. delavayi and putative hybrids
编号 Code | 类型 Type | 差异位点 Variable sites (bp) | ||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 128 | 135 | 211 | 214 | 245 | 254 | 254 | 299 | 313 | 333 | 337 | 340 | 344 | 348 | 351 | 358 | 365-369 | ||
BLD | D1 | G | A | T | C | G | C | G | A | C | A | A | G | T | G | G | T | T | T | AATTC |
SBSD | D2 | - | G | * | * | * | * | * | * | * | * | * | A | A | A | A | A | G | G | ***** |
BLM | D3 | * | * | A | G | A | C | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
SBSM | D4 | * | * | A | G | A | A | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
AH | D1 | G | A | T | C | G | C | G | A | C | A | A | G | T | G | G | T | T | T | AATTC |
SBSH5 | D2 | * | G | * | * | * | * | * | * | * | * | * | A | A | A | A | A | G | G | ***** |
BLH | D3 | * | * | A | G | A | C | T | T | T | T | G | * | * | * | * | * | * | * | G---- |
SBSH2/3/4/7/8/9 | D4 | * | * | A | G | A | A | T | T | T | T | G | * | * | * | * | * | * | · | G---- |
图1 大白杜鹃、马缨杜鹃及疑似杂交后代的联合ITS、trnL-trnF、rbcL、psbA-trnH序列片段的聚类图。AH、BLH、SBSH、BLM、BLD、SBSM和SBSD含义见表1。
Fig. 1 Clustering chart of combined ITS, trnL-trnF, rbcL, psbA-trnH sequence of Rhododendron decorum, R. delavayi and putative hybrids. The means of AH, BLH, SBSH, BLM, BLD, SBSM, and SBSD see Table 1.
图3 大白杜鹃、马缨杜鹃、疑似杂交后代与人工杂交后代部分数量性状。AH、BLH和SBSH含义见表1。
Fig. 3 Part of quantitative characters of Rhododendron decorum, R. delavayi, putative and artificial hybrid. The means of AH, BLH, and SBSH see Table 1.
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